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Literature summary extracted from
- A 4-methoxybenzoate O-demethylase from Pseudomonas putida. A new type of monooxygenase system (1975), Eur. J. Biochem., 57, 241-256.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.99.15 | Putidamonooxin | - |
Pseudomonas putida |
General Stability
| EC Number | General Stability | Organism |
|---|---|---|
| 1.14.99.15 | ethanol, 5-15% v/v, stabilizes activity in buffer and crude extract | Pseudomonas putida |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.99.15 | 1,10-phenanthroline | - |
Pseudomonas putida |  |
| 1.14.99.15 | 2,2'-dipyridyl | - |
Pseudomonas putida | |
| 1.14.99.15 | 5,5'-dithiobis(2-nitrobenzoate) | - |
Pseudomonas putida | |
| 1.14.99.15 | 8-hydroxyquinoline | - |
Pseudomonas putida | |
| 1.14.99.15 | Amytal | - |
Pseudomonas putida | |
| 1.14.99.15 | Atebrin | - |
Pseudomonas putida | |
| 1.14.99.15 | Bathocuproinedisulfonate | - |
Pseudomonas putida | |
| 1.14.99.15 | bathophenanthrolinedisulfate | - |
Pseudomonas putida | |
| 1.14.99.15 | diethyldithiocarbamate | - |
Pseudomonas putida | |
| 1.14.99.15 | EDTA | - |
Pseudomonas putida | |
| 1.14.99.15 | KCN | - |
Pseudomonas putida | |
| 1.14.99.15 | p-chloromercuribenzoate | reversible by GSH | Pseudomonas putida | |
| 1.14.99.15 | rotenone | - |
Pseudomonas putida | |
| 1.14.99.15 | Thenoyl trifluoroacetone | - |
Pseudomonas putida | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.99.15 | 0.0007 | - |
3-Nitro-4-methoxybenzoate | - |
Pseudomonas putida | |
| 1.14.99.15 | 0.0014 | - |
4-Methoxybenzoate | - |
Pseudomonas putida | |
| 1.14.99.15 | 0.008 | - |
NADH | reductase | Pseudomonas putida | |
| 1.14.99.15 | 0.077 | - |
N-methyl-4-aminobenzoate | - |
Pseudomonas putida | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.99.15 | Fe2+ | both enzyme components are iron-sulfur proteins | Pseudomonas putida | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.99.15 | 4-methoxybenzoate + NADH + O2 | Pseudomonas putida | - |
4-hydroxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.99.15 | Pseudomonas putida | - |
- |
- |
Oxidation Stability
| EC Number | Oxidation Stability | Organism |
|---|---|---|
| 1.14.99.15 | putidamonooxin and NADH-reductase, extremely O2-sensitive, purification and storage of the reductase under anaerobic conditions | Pseudomonas putida |
Posttranslational Modification
| EC Number | Posttranslational Modification | Comment | Organism |
|---|---|---|---|
| 1.14.99.15 | glycoprotein | 30% carbohydrate in each of the 50000 Da subunits | Pseudomonas putida |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.99.15 | - |
Pseudomonas putida |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.14.99.15 | 10.97 | - |
- |
Pseudomonas putida |
| 1.14.99.15 | 21.96 | - |
4-methoxybenzoate | Pseudomonas putida |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.14.99.15 | -20°C, freeze-dried partially purified reductase stable for months without loss of activity, purified reductase is 3-4 weeks stable after addition of NADH in N2-atmosphere | Pseudomonas putida |
| 1.14.99.15 | 4°C, concentrated putidamonooxin stable for several months in N2-atmosphere with DTT and dithionite | Pseudomonas putida |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.99.15 | 3-nitro-4-methoxybenzoate + NADH + O2 | - |
Pseudomonas putida | 3-nitro-4-hydroxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
| 1.14.99.15 | 4-methoxybenzoate + NADH + O2 | - |
Pseudomonas putida | 4-hydroxybenzoate + NAD+ + H2O + formaldehyde | - |
? | |
| 1.14.99.15 | N-methyl-4-aminobenzoate + NADH + O2 | - |
Pseudomonas putida | 4-aminobenzoate + NAD+ + H2O + formaldehyde | - |
? | |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.99.15 | 30 | - |
assay at | Pseudomonas putida |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.99.15 | 8 | - |
- |
Pseudomonas putida |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.99.15 | 7.3 | 9 | about half-maximal activity at pH 7.3 and 9.0, reductase | Pseudomonas putida |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.99.15 | FMN | - |
Pseudomonas putida | |
| 1.14.99.15 | NADH | - |
Pseudomonas putida | |
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